Amino and nitro containing tricyclic compounds useful as inhibitors of ace

ABSTRACT

The present invention relates to compounds of the formula ##STR1## wherein A is methylene, oxygen, sulfur or N--B wherein B is R 1  or COR 2  wherein R 1  is hydrogen, a C 1  -C 4  alkyl or an Ar--Z-- group wherein Ar is aryl and Z is a C 0  -C 4  alkyl and R 2  is a --CF 3 , a C 1  -C 10  alkyl or an Ar--Z group; 
     R is hydrogen or a C 1  -C 4  alkyl; and 
     X and Y are each independently hydrogen, nitro or amino, with the proviso that when X is nitro or amino, Y must be hydrogen, and when Y is nitro or amino, X must be hydrogen; and 
     pharmaceutically acceptable salts thereof, which are inhibitors of Angiotensin Converting Enzyme.

This is a division of application Ser. No. 08/046,326, filed Apr. 12,1993, U.S. Pat. No. 5,308,841, which is a divisional of application Ser.No. 07/979,029, filed Nov. 20, 1992, now abandoned, which is adivisional of application Ser. No. 07/905,494, filed Jun. 25, 1992, nowU.S. Pat. No. 5,208,230, issued May 4, 1993, which is a continuation ofapplication Ser. No. 07/777,625, filed Oct. 23, 1991, now abandoned,which is a continuation-in-part of application Ser. No. 07/633,572,filed Dec. 21, 1990, now abandoned.

BACKGROUND OF THE INVENTION

The compounds of the present invention are inhibitors ofAngiotensin-Converting Enzyme (ACE). ACE is a peptidyl dipeptidase whichcatalyzes the conversion of angiotensin I to angiotensin II. AngiotensinII is a powerful vasopressor which also stimulates aldosterone secretionby the adrenal cortex.

Inhibition of ACE lowers levels of angiotensin II and thus inhibits thevasopressor, hypertensive and hyperaldosteronemic effects causedthereby. It is known that inhibition of ACE is useful in the treatmentof patients suffering from disease states such as hypertension andchronic congestive heart failure [See William W. Douglas,"Polypeptides--Angiotensin, Plasma Kinins, and Others", Chapter 27, inGOODMAN AND GILLMAN'S THE PHARMACOLOGICAL BASIS OF THERAPEUTICS, 7thedition, 1985, pp. 652-3, MacMillan Publishing Co., New York, N.Y.]. Inaddition, it has been disclosed that ACE inhibitors are useful intreating cognitive disorders [German Application No. 3901-291-A,published Aug. 3, 1989].

Certain tricyclic compounds such as that described by Flynn et al. [J.Amer. Chem. Soc. 109, 7914-15 (1987)] and Flynn and Beight [EuropeanPatent Application Publication No. 0 249 223 A2, published Dec. 16,1987] are known as ACE inhibitors. The compounds of the presentinvention differ from these tricyclic inhibitors of ACE by having anamino or nitro substituent on the aromatic moiety of the tricyclicstructure.

It has now been found that the amino or nitro substituted compounds ofthe present invention possess an unexpectedly prolonged duration ofactivity in comparison to other ACE inhibitors of similar structure.

SUMMARY OF THE INVENTION

The present invention provides novel compounds of the formula (1)##STR2## wherein A is methylene, oxygen, sulfur or N--B wherein B is R₁or COR₂ wherein R₁ is hydrogen, a C₁ -C₄ alkyl or an Ar--Z-- groupwherein Ar is aryl and Z is a C₀ -C₄ alkyl and R₂ is

a --CF₃, a C₁ -C₁₀ alkyl or an Ar--Z group;

R is hydrogen or a C₁ -C₄ alkyl; and

X and Y are each independently hydrogen, nitro or amino, with theproviso that one of X and Y is hydrogen and one of X and Y is other thanhydrogen; and

the pharmaceutically acceptable salts thereof.

The present invention further provides a method of inhibiting ACE in apatient in need thereof comprising administering to said patient aneffective ACE inhibitory amount of a compound of formula (1).

In addition, the present invention provides a composition comprising anassayable amount of a compound of formula (1) in admixture or otherwisein association with an inert carrier. The present invention alsoprovides a pharmaceutical composition comprising an effectiveimmunosuppressive amount of a compound of formula (1) in admixture orotherwise in association with one or more pharmaceutically acceptablecarriers or excipients.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the term "C₁ -C₄ alkyl" refers to a saturated straightor branched chain hydrocarbyl radical of one to four carbon atoms andincludes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tertiarybutyl and the like. The term "C₁ -C₁₀ alkyl" refers to a saturatedstraight or branched chain hydrocarbyl radical of one to ten carbonatoms, respectively, including methyl, ethyl, propyl, isopropyl,n-butyl, isobutyl, tertiary butyl, pentyl, isopentyl, hexyl,2,3-dimethyl-2-butyl, heptyl, 2,2-dimethyl-3-pentyl, 2-methyl-2-hexyl,octyl, 4-methyl-3-heptyl and the like. The term "halogen", "halo","halide" or "X" refers to a chlorine, bromine, or iodine atom.

As used herein, the term "Ar--Z--" refers to a radical wherein Ar is anaryl group and Z is a C₀ -C₄ alkyl. The term "Ar" refers to a phenyl ornaphthyl group unsubstituted or substituted with from one to threesubstituents selected from the group consisting of methylenedioxy,hydroxy, C₀ -C₄ alkoxy, fluoro and chloro. The term "C₀ -C₄ alkyl"refers to a saturated straight or branched chain hydrocarbyl radical ofzero to four carbon atoms and includes a bond, methyl, ethyl, propyl,isopropyl, n-butyl, isobutyl, tertiary butyl and the like. Specificallyincluded within the scope of the term "Ar--Z--" are phenyl, naphthyl,phenylmethyl or benzyl, phenylethyl, p-methoxybenzyl, p-fluorobenzyl andp-chlorobenzyl.

The compounds of formula (1), wherein R is hydrogen can be prepared byutilizing procedures and techniques well known and appreciated by one ofordinary skill in the art. A general synthetic scheme for preparingthese compounds is set forth in Scheme A wherein all substituents,unless otherwise indicated, are previously defined. ##STR3## Scheme Aprovides a general synthetic scheme for preparing compounds of formula(1) wherein R is hydrogen.

In step a, the appropriate phthalimide protected amino/carboxylic acidcompound of structure (1) is nitrated to give a mixture of thecorresponding 9-nitro and 11-nitro/phthalimide protectedamino/carboxylic acid compounds of structure (2).

For example, the appropriate phthalimide protected amino/carboxylic acidcompound of structure (1) is contacted with a molar excess of anitrating agent, such as nitronium tetrafluoroborate. The reactants aretypically contacted in a suitable organic solvent, such as methylenechloride. The reactants are typically stirred together for a period oftime ranging from 10-50 hours and at a temperature range of from -60° C.to 10° C. The 9-nitro and 11-nitro/phthalimide protectedamino/carboxylic acid compounds of structure (2) is recovered from thereaction mixture by extractive methods as is known in the art. They canbe separated by silica gel chromatography.

In step b, the appropriate individual 9-nitro and 11-nitro/phthalimideprotected amino/carboxylic acid compounds of structure (2) is esterifiedto give the corresponding individual 9-nitro and 11-nitro/phthalimideprotected amino/carboxylic acid, diphenylmethyl ester compounds ofstructure (3).

For example, the appropriate individual 9-nitro and 11-nitro/phthalimideprotected amino/carboxylic acid compounds of structure (2) is contactedwith a molar equivalent of an appropriate diphenylmethylating agent,such as diphenyldiazomethane. The reactants are typically contacted in asuitable organic solvent, such as methylene chloride. The reactants aretypically stirred together at room temperature for a period of timeranging from 1-10 days. The corresponding individual 9-nitro and11-nitro/phthalimide protected amino/carboxylic acid, diphenylmethylester compounds of structure (3) is recovered from the reaction zone byevaporation of the solvent. They may be purified by silica gelchromatography.

In step c, the phthalimide protecting group of the appropriateindividual 9-nitro and 11-nitro/phthalimide protected amino/carboxylicacid, diphenylmethyl ester compounds of structure (3) is removed to givethe corresponding individual 9-nitro and 11-nitro/amino/carboxylic acid,diphenylmethyl ester compounds of structure (4).

For example, the appropriate individual 9-nitro and 11-nitro/phthalimideprotected amino/carboxylic acid, diphenylmethyl ester compounds ofstructure (3) is contacted with a molar excess of hydrazine monohydrate.The reactants are typically contacted in protic organic solvent, such asmethanol. The reactants are typically stirred together at roomtemperature for a period of time ranging from 5-24 hours. Thecorresponding individual 9-nitro and 11-nitro/amino/carboxylic acid,diphenylmethyl ester compounds structure (4) is recovered from thereaction zone by evaporation of the solvent, redissolving in CHCl₃,filtration to remove phthal-hydrazide and removal of the CHCl₃ in vacuo.

In step d, the amino functionality of the appropriate individual 9-nitroand 11-nitro/amino/carboxylic acid, diphenylmethyl ester compound s ofstructure (4) is alkylated with(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid, diphenylmethylester (5) to give the corresponding individual 9-nitro and11-nitro/[1-(carbodiphenylmethoxy)-3-phenyl]propylamino/carboxylic acidcompounds, diphenylmethyl ester compounds of structure (6).

For example, the appropriate individual 9-nitro and11-nitro/amino/carboxylic acid, diphenylmethyl ester compounds ofstructure (4) is contacted with a molar excess of(R)-trifluoromethanesulfonyl-4-phenylbutyric acid, diphenylmetheyl ester(5) and a molar excess of strong, non-nucleophilic base, such as ProtonSponge. The reactants are typically contacted in a suitable organicsolvent such as methylene chloride. The reactants are typically stirredtogether at room temperature for a period of time ranging from 10-30hours. The corresponding individual 9-nitro and11-nitro/[1-(carbodiphenylmethoxy)-3-phenyl]propylamino/carboxylic acid,diphenylmethyl ester compounds of structure (6) is recovered from thereaction zone by silica gel chromatography.

In step e₁, the diphenylmethyl ester functionalities of the appropriateindividual 9-nitro and11-nitro/[1-(carbodiphenylmethoxy)-3-phenyl]propylamino/carboxylic acid,diphenylmethyl ester compounds of structure (6) is hydrolyzed to givethe corresponding individual 9-nitro and11-nitro/[1-(carboxy)-3-phenyl]propylamino/carboxylic acid compounds ofstructure (7).

For example, the appropriate individual 9-nitro and11-nitro/[1-(carbodiphenylmethoxy)-3-phenyl]propylamino/carboxylic acid,diphenylmethyl ester compounds of structure (6) is contacted with amolar excess of an acid, such as trifluoroacetic acid. The reactants aretypically contacted in a suitable organic solvent such as methylenechloride. The reactants are typically stirred together at roomtemperature for a period of time ranging from 1-24 hours. Thecorresponding individual 9-nitro and11-nitro/[1-(carboxy)-3-phenyl]propylamino/carboxylic acid compounds ofstructure (7) is recovered from the reaction zone by removal of solventand trituration to remove nonpolar by-products followed by reverse phaseHPLC where required.

In step e₂, the nitro functionality of the appropriate individual9-nitro and 11-nitro/[1-(carboxy)-3-phenyl]propylamino/carboxylic acid,diphenylmethyl ester compounds of structure (6) is reduced concurrentlywith diphenylmethyl ester hydrogenolysis to give the correspondingindividual 9-amino and11-amino/[1-(carboxy)-3-phenyl]propylamino/carboxylic acid compounds ofstructure (8).

For example, the individual 9-nitro and11-nitro/[1-(carboxy)-3-phenyl]propylamino/carboxylic acid,diphenylmethyl ester compounds of structure (6) is contacted with acatalytic amount of a hydrogenation catalyst, such as 10%palladium/carbon. The reactants are typically contacted in a suitablesolvent mixture such as tetrahydrofuran/water. The reactants aretypically shaken under a hydrogen atmosphere of 35-45 psi at roomtemperature for a period of time ranging from 5-24 hours. The individual9-amino and 11-amino/[1-(carboxy)-3-phenyl]propylamino/carboxylic acidcompounds of structure (8) is recovered from the reaction zone byevaporation of the solvent. They may then be converted to theirdihydrochloride salts and triturated with hexane to remove thediphenylmethane.

For those compounds of formula (1) wherein A is N--COR₂, the N--COR₂group of the appropriate individual 9-nitro and11-nitro/[1-(carboxy)-3-phenyl]propylamino/carboxylic acid compounds ofstructure (7) or the appropriate individual 9-amino and11-amino/[1-(carboxy)-3-phenyl]propylamino/carboxylic acid compounds ofstructure (8) may be cleaved by techniques and procedures well known anappreciated in the art, such as lithium hydroxide, to give thecorresponding compounds of formula (1) wherein A is NH.

The compounds of formula (1), wherein R is a C₁ -C₄ alkyl can beprepared by utilizing procedures and techniques well known andappreciated by one of ordinary skill in the art. A general syntheticscheme for preparing these compounds is set forth in Scheme B whereinall substituents, unless otherwise indicated, are previously defined.##STR4##

Scheme B provides a general synthetic scheme for preparing compounds offormula (1) wherein R is C₁ -C₄ alkyl.

In step a, the amino functionality of the appropriate individual 9-nitroand 11-nitro/amino/carboxylic acid, diphenylmethyl ester compounds ofstructure (4) is alkylated with(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid, C₁ -C₄ alkyl ester(9) to give the corresponding individual 9-nitro and11-nitro/[1-(carboalkoxy)-3-phenyl]propylamino/carboxylic acid,diphenylmethyl ester compounds of structure (10) as described previouslyin Scheme A, step d.

In step b₁, the carboxylic acid, diphenylmethyl ester functionality ofthe appropriate individual 9-nitro and11-nitro/[1-(carboalkoxy)-3-phenyl]propylamino/carboxylic acid,diphenylmethyl ester compounds of structure (10) is hydrolyzed to givethe corresponding individual 9-nitro and11-nitro/[1-(carboalkoxy)-3-phenyl]propylamino/carboxylic acid compoundsof structure (11) as described previously in Scheme A, step e₁.

In step b₂, the nitro functionality of the appropriate individual9-nitro and 11-nitro/[1-(carboalkoxy)-3-phenyl]propylamino/carboxylicacid, diphenylmethyl ester compounds of structure (10) is reducedconcurrently with diphenylmethyl ester hydrogenolysis to give thecorresponding individual 9-amino and11-amino/[1-(carboalkoxy)-3-phenyl]propylamino/carboxylic acid compoundsstructure (12) as described previously in Scheme A, step e₂.

For those compounds of formula (1) wherein A is N--COR₂, the N--COR₂group of the appropriate individual 9-nitro and11-nitro/[1-(carboalkoxy)-3-phenyl]propylamino/carboxylic acid compoundsof structure (11) or the appropriate 9-amino and11-amino/[1-(carboalkoxy)-3-phenyl]propylamino/carboxylic acid compoundsstructure (12) may be cleaved by techniques and procedures well known anappreciated in the art, such as lithium hydroxide, to give thecorresponding compounds of formula (1) wherein A is NH.

Phthalimide protected amino/carboxylic acid compounds of structure 1wherein A is 0 may be prepared as described in Scheme C. In Scheme C,all substituents unless otherwise indicated are as previously defined.##STR5##

Scheme C provides a general synthetic procedure for preparingphthalimide protected amino/carboxylic acid compounds of structure 1wherein A is 0.

In step a, the appropriate phthalimide blocked (S)-phenylalaninederivative of structure (13) is converted to the corresponding acidchloride, then reacted with the appropriate L-serine methyl ester ofstructure (14) to give the corresponding 1-oxo-3-phenylpropyl-L-serinemethyl ester of structure (15).

For example, the appropriate phthalimide blocked (S)-phenylalaninederivative of structure (13) can be reacted with oxalyl chloride in asuitable aprotic solvent, such as methylene chloride. The resulting acidchloride can then be coupled with the appropriate L-serine methyl esterof structure (14) using N-methylmorpholine in a suitable aproticsolvent, such as dimethylformamide, to give the appropriate1-oxo-3-phenylpropyl-L-serine methyl ester of structure (15).

In step b, the hydroxy functionality of the appropriate1-oxo-3-phenylpropyl-L-serine methyl ester of structure (15) isallylated with the allyl imidate of structure (16) to give thecorresponding 1-oxo-3-phenylpropyl-L-serine-O-allyl methyl ester ofstructure (17).

For example, the appropriate 1-oxo-3-phenylpropyl-L-serine methyl esterof structure (15) is contacted with 2 molar equivalents of the allylimidate of structure (16) and a molar equivalent of a suitable acid suchas trifluoromethanesulfonic acid. The reactants are typically contactedin a suitable organic solvent mixture such as methylenechloride/cyclohexane. The reactants are typically stirred together atroom temperature under an inert atmosphere for a period of time rangingfrom 2-24 hours. The 1-oxo-3-phenylpropyl-L-serine-O-allyl methyl esterof structure (17) is recovered from the reaction zone by extractivemethods as is known in the art. It may be purified by silica gelchromatography or crystalization.

In step c, the appropriate 1-oxo-3-phenylpropyl-L-serine-O-allyl methylester of structure (17) is cyclized to give the corresponding(4S)-enamine of structure (18).

For example, the appropriate 1-oxo-3-phenylpropyl-L-serine-O-allylmethyl ester of structure (17) is first contacted with a molar excess ofa mixture of ozone/oxygen. The reactants are typically contacted in asuitable organic solvent mixture such as methylene chloride/methanol.The reactants are typically stirred together for a period of timeranging from 5 minutes to 30 minutes or until a blue color persists andat a temperature range of from -78° C. to -40° C. The reaction isquenched with an excess of methylsulfide and the intermediate aldehydecompound recovered from the reaction zone by extractive methods as isknown in the art.

The intermediate aldehyde compound is then contacted withtrifluoroacetic acid in a suitable aprotic solvent, such as methylenechloride to give the corresponding (4S)-enamine of structure (18).

In step d, the appropriate (4S)-enamine of structure (18) is cyclizedand the methyl ester functionality is removed to give the correspondingphthalimide protected amino/carboxylic acid compound of structure (19)by an acid catalyzed Friedel-Crafts reaction. For example, theappropriate (4S)-enamine of structure (18) can be converted to thecorresponding phthalimide protected amino/carboxylic acid compound ofstructure (19) by treatment with a mixture of trifluoromethane sulfonicacid and trifluoroacetic anhydride in a suitable aprotic solvent, suchas methylene chloride.

Phthalimide protected amino/carboxylic acid compounds of structure 1wherein A is N--B may be prepared as described in Scheme D. In Scheme D,all substituents unless otherwise indicated are as previously defined.##STR6##

Scheme D provides an alternative general synthetic procedure forpreparing phthalimide protected amino/carboxylic acid compounds ofstructure 1 wherein A is N--B.

In step a, the appropriate phthalimide blocked (S)-phenylalaninederivative of structure (13) is converted to the corresponding acidchloride, then reacted with the appropriate3-trifluoracetylamino-3-allyl-L-2-aminopropionic acid, methyl ester ofstructure (21) to give the corresponding1-oxo-3-phenylpropyl-N-trifluoracetyl-N-allyl-L-amino acid, methyl esterof structure (22) as described previously in Scheme C, step a.

In step b, the appropriate1-oxo-3-phenylpropyl-N-trifluoracetyl-N-allyl-L-amino acid methyl esterof structure (22) is cyclized to give the corresponding enamine ofstructure (23) as described previously in Scheme C, step c.

In step c, the appropriate (4S)-enamine of structure (23) is cyclized togive the corresponding phthalimide protected amino/carboxylic acidcompound of structure (24) as described previously in Scheme C, step d.

In optional step d, it is necessary to reesterify the carboxyfunctionality of the phthalimide protected amino/carboxylic acidcompound of structure (24) in order to carry out optional steps e and f.For example, treatment of the crude phthalimide protectedamino/carboxylic acid compound of structure (24) withbromodiphenylmethane in a suitable aprotic solvent, such asdimethylformamide along with a non-nucleophilic base, such as cesiumcarbonate, may be used to give the corresponding phthalimide protectedamino/carboxylic acid, diphenylmethyl ester compound of structure (25).

In optional step e, the trifluroacetate protecting group of theappropriate phthalimide protected amino/carboxylic acid, diphenylmethylester compound of structure (25) is removed with a base such as lithiumhydroxide as is known in the art to give the corresponding phthalimideprotected amino/carboxylic acid, diphenylmethyl ester compound ofstructure (26).

In optional step f, the free amino functionality of the phthalimideprotected amino/carboxylic acid, diphenylmethyl ester compound ofstructure (26) is alkylated or acylated by techniques and proceduresknown in the art to give the corresponding phthalimide protectedamino/carboxylic acid, diphenylmethyl ester compound of structure (27)wherein B is R₁ or COR₂ which may be used in Scheme A.

In step g, the diphenylmethyl ester functionality of the appropriatephthalimide protected amino/carboxylic acid, diphenylmethyl estercompound of structure (25), the appropriate phthalimide protectedamino/carboxylic acid, diphenylmethyl ester compound of structure (26)or the appropriate phthalimide protected amino/carboxylic acid,diphenylmethyl ester compound of structure (27) is removed to give thecorresponding phthalimide protected amino/carboxylic acid compound ofstructure (28) as described previously in Scheme A, step e₁.

Starting materials for use in Schemes A through D are readily availableto one of ordinary skill in the art. For example, certain tricyclicamino compounds of structure 1 wherein X is S are described in EuropeanPatent 0 249 223 (Dec. 16, 1987) and certain other tricyclic aminocompounds of structure 1 wherein A is methylene may be prepared asdescribed in European Patent Application of Flynn and Beight,Application #34533A EP (Jun. 11, 1987). Diphenyldiazomethane isdescribed in Org. Syn., Coll. Vol. (III), 351. Certain(R)-2-hydroxy-4-phenylbutyric acid esters are described in U.S. Pat. No.4,837,354 of Flynn and Beight (Jun. 6, 1989). Ethyl(R)-2-trifluoromethanesulfonyl-4-phenylbutyrate is described inTetrahedron Lett. 25, 1143-46 1984.

The following examples present typical syntheses as described in SchemesA through D. These examples are understood to be illustrative only andare not intended to limit the scope of the present invention in any way.As used herein, the following terms have the indicated meanings: "g"refers to grams; "mmol" refers to millimoles; "mL" refers tomilliliters; "bp" refers to boiling point; "°C." refers to degreesCelsius; "mm Hg" refers to millimeters of mercury; "μL" refers tomicroliters; "μg" refers to micrograms; and "μM" refers to micromolar.

EXAMPLE 1[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2 ]benzazepine-4-carboxylic acid

Scheme A, Step a:[4S-[4α,7α(R*),12bβ]]-7-[1,3,-Dihydro1,3-dioxo-2H-isoindol-2-yl]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid and[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-1,2,3,4,6,7,8,12b-octahydro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid (1.46 g, 3.5 mmol) in methylene chloride (10 mL) and cool to -60°C. Add, by dropwise addition, a solution of nitronium tetrafluoroborate(25 mL of a 0.5M solution in sulfolane, 12.5 mmol) in methylene chloride(15 mL). Warm slowly to 10° C. over 34 hours. Partition betweenmethylene chloride (75 mL) and water (75 mL). Separate the aqueous phaseand extract with methylene chloride (50 mL). Combine the organic phases,dry (MgSO₄) and evaporate the solvent in vacuo. Purify and separate byflash silica gel chromatography (1:1 ethyl acetate/hexane→1:1 ethylacetate/hexane with 5% acetic acid→2:1 ethyl acetate/hexane with 5%acetic acid) to give the 11-nitro title compound (1.01 g, 64.3%) and the9-nitro title compound (0.43 g, 27.4%) as brown foams.

Scheme A, Step b:[4S-[4α,7α(R*),12bβ]]-7-[1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl[-1,2,3,4,6,7,8,12b,-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid (674 mg, 1.50 mmol) in methylene chloride (1 mL). Adddiphenyldiazomethane (291 mg, 1.50 mol) and let stand for several daysin which time the methylene chloride evaporates. Purify the residue bysilica gel chromatography (40% ethyl acetate/hexane→55% ethylacetate/hexane) to give the title compound as a white foam (613 mg,66%).

Anal. Calcd for C₃₆ H₂₉ N₃ O₇ : C, 70.23; H, 4,75; N, 6.83; Found: C,70.23; H, 4,77; N, 6.63.

Scheme A, Step c:[4S-[4α,7α(R*),12bβ]]-7-Amino-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Slurry[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (200 mg, 0.324 mmol) in methanol (6 mL) andadd hydrazine hydrate (1.0 mL of a 1M solution in methanol, 1.0 mmol).Stir at room temperature under an argon atmosphere for 20 hours. Dilutewith methylene chloride (30 mL) and stir for 2 hours. Filter andconcentrate in vacuo. Take up the residue in methylene chloride andfilter again. Wash the filtrate with water, dry (MgSO₄) and evaporatethe solvent in vacuo to give the title compound as a yellow foam (165mg).

Scheme A, Step d:[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[Carbodiphenylmethoxy]-3-phenylpropyl]amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Dissolve (R)-2-hydroxy-4-phenylbutyric acid, ethyl ester (4.16 g, 20mmol) in methanol (40 mL). Add 1N lithium hydroxide (25 mL, 25 mmol) andstir under an atmosphere of argon for 3.5 hours. Acidify with 1Nhydrochloric acid (25 mL), saturate with sodium chloride and extractwith ethyl acetate (3×25 mL). Combine the organic phases, dry (MgSO₄)and evaporate the solvent in vacuo. Take up the residue in methylenechloride (20 mL) and add diphenyldiazomethane (3.36 g, 17.3 mmol). Stirovernight and wash with saturated sodium hydrogen carbonate (2×). Dry(MgSO₄), evaporate the solvent in vacuo and recrystallize (methylenechloride/hexane then again with cyclohexane) to give(R)-2-hydroxy-4-phenylbutyric acid, diphenylmethyl ester as whiteneedles (2.3 g, 38%); mp 82°-84° C.

Anal. Calcd for C₂₃ H₂₂ O₃ : C, 79.74; H, 6.40; Found: C, 79,76; H,6.41.

Dissolve (R)-2-hydroxy-4-phenylbutyric acid, diphenylmethyl ester (1.38g, 4.0 mmol) in methylene chloride (12 mL). Add pyridine (0.32 mL, 4mmol), place under an argon atmosphere and cool to -10° C. Add, bydropwise addition, a solution of trifluoromethanesulfonic anhydride(0.673 mL, 4.0 mmol) in methylene chloride (2 mL). Stir at -10° to -20°C. for 2 hours, dilute with hexane (10 mL) and filter. Evaporate thesolvent in vacuo to give (R)-2-trifluoromethanesulfonyl-4-phenylbutyricacid, diphenylmethyl ester.

Dissolve [4S-[4α,7α(R*),12bβ]]-7-amino-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (158 mg, 0.632 mmol) and proton sponge (138mg, 0. 648 mmol) in methylene chloride (5 mL). Add a solution of(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid , diphenylmethylester (309 mg, 0.648 mmol) in methylene chloride (2 ml ). Stir at roomtemperature under an argon atmosphere for 26 hours. Purify by flashsilica gel chromatography (30% ethyl acetate/hexane→50% ethylacetate/hexane) to give the title compound as a yellow foam (178 mg,67.2%).

Anal. Calcd for C₅₁ H₄₇ N₃ O₇ : C, 75.25; H, 5,82; N, 5.16; Found: C,74.10; H, 5,82; N, 4.87.

Scheme A, Step e₁ :[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2]benzazepine-4-carboxylicacid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[carbodiphenylmethoxy]-3-phenylpropyl]amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (39 mg, 0.045 mmol) in methylene chloride (1mL). Add trifluoroacetic acid (1 mL) and anisole (0.2 mL). Allow tostand for 3 hours under an argon atmosphere. Partition between methylenechloride and 1N hydrochloric acid. Separate the aqueous phase and freezedry. Purify by High Pressure Liquid Chromatography (elute with 25%methanol with 0.1% trifluoroacetic acid at 15 mL/min on a 22.5 cm×250 cmVydac C-18 column) to give the title compound (18 mg, 72%).

EXAMPLE 2[4S-[4α,7α(R*),12bβ]]-9-Amino-7-[(S-1-carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, dihydrochloride

Dissolve[4S-[4α7α(R*),12bβ]]-7-[[S-1-[carbodiphenylmethoxy]-3-phenylpropyl]amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (52 mg, 0.064 mmol) in tetrahydrofuran (3mL). Add 10% palladium/carbon (18 mg) and water (3 mL). Shake under 44psi of hydrogen at room temperature for 20 hours. Filter and evaporatethe solvent in vacuo. Take the residue up in 1N hydrochloric acid,filter then freeze dry to give the title compound (25 mg, 74%).

EXAMPLE 3[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid

Scheme A, Step b:[4S-[4α,7α(R*),12bβ]]-7-[1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

dissolve[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-1,2,3,4,6,7,8,12,b-octahydro-11-nitro-6-oxopyrido[2,1-a]benzazepine-4-carboxylicacid (795 mg, 1.77 mmol) in methylene chloride (2 mL). Adddiphenyldiazomethane (350 mg, 1.78 mmol) and methylene chloride (5 mL).Let stand for 8 hours. Purify the residue by silica gel chromatography(35% ethyl acetate/hexane→50% ethyl acetate/hexane) to give the titlecompound as a white foam (756 mg, 69.4%).

Scheme A, Step c:[4S-[4α,7α(R*),12bβ]]-7-Amino-1,2,3,4,5,6,7,8,12b-octahydro-11-nitro6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Slurry[4S-[4α,7α(R*),12bβ]]-7-dihydro-1,3-dioxo-2H-isoindol-2-yl]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (756 mg, 1.23 mmol) in methanol (5 mL) andadd hydrazine hydrate (4.0 mL of a 1M solution in methanol, 4.0 mmol).Stir at room temperature under an argon atmosphere for 10 minutes thenwarm to reflux for 90 minutes. Evaporate the solvent in vacuo and stirthe residue with methylene chloride for 20 minutes. Filter and wash thefiltrate with water, dry (MgSO₄) and evaporate the solvent in vacuo togive the title compound as a yellow film (610 mg, 00%).

Scheme A, Step b:[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[Carbodiphenylmethoxy]-3-phenylpropyl]amino]-1,2,3,4,6,6,8,,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-amino-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2-benzazepine-4-carboxylicacid, diphenylmethyl ester (90 mg, 0.18 mmol) and proton sponge (250 mg,1.12 mmol) in methylene chloride (5 mL). Add a solution of(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid, diphenylmethylester (520 mg, 1.17 mmol) in methylene chloride (6 ml). Stir at roomtemperature under an argon atmosphere for 22 hours. Dilute withcyclohexane (20 mL) and filter. Purify by silica gel chromatography (30%ethyl acetate/hexane) to give the title compound (105 mg, 72%).

Anal. Calcd for C₅₁ H₄₇ N₃ O₇ : C, 75.25; H, 5.82; N, 5.16; Found: C,74.84; H, 5.84; N, 4.84.

Scheme A, Step e₁ :[bS-[4α,7α(R*),12bβ]]-7-(S-1-Carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, hydrochloride

dissolve[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[carbodiphenylmethoxy]-3-phenylpropyl]amino]-1,2,3,4,6,7,812b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (52 mg, 0.063 mmol) in methylene chloride (2mL). Add trifluoroacetic acid (1 mL) and anisole (0.2 mL). Allow tostand for 2 hours under an argon atmosphere. Partition between methylenechloride and 9:1 water/methanol. Separate the organic phase and extractwith additional 9:1 water/methanol (10 mL). Combine the aqueous phasesand wash with methylene chloride (3×10 mL). Separate the aqueous phaseand freeze dry to give the title compound as a white powder (24.6 mg,74.5%).

EXAMPLE 4[4S-[4α,7α(R*),12bβ]]-11-Amino-7-[(S-1-carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-6-oxopyrido[2,1-][2]benzazepine-4-carboxylicacid, dihydrochloride

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[carbodiphenylmethoxy]-3-phenylpropyl]amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (56 mg, 0.06 mmol) in tetrahydrofuran (6 mL).Add 10% palladium/carbon (20 mg) and water (3 mL). Shake under 45 psi ofhydrogen at room temperature for 5 hours. Filter and partition thefiltrate between in 1N hydrochloric acid and methylene chloride.Separate the aqueous phase and freeze dry to give the title compound(35.3 mg, 98%).

EXAMPLE 5[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid

Scheme B, Step a:[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-amino-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (596 mg, 1.23 mmol) and proton sponge (296mg, 1.25 mmol) in methylene chloride (10 mL). Add a solution of(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid, ethyl ester (428mg, 1.25 mmol) in methylene chloride (6 ml). Stir at room temperatureunder an argon atmosphere overnight. Purify by silica gel chromatographyto give the title compound.

Scheme B, Step b₁ :[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[(S-1-carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (42.5 mg, 0.063 mmol) in methylene chloride(2 mL). Add trifluoroacetic acid (1 mL) and anisole (0.2 mL). Allow tostand for 2 hours under an argon atmosphere. Partition between methylenechloride and 9:1 water/methanol. Separate the organic phase and extractwith additional 9:1 water/methanol (10 mL). Combine the aqueous phasesand wash with methylene chloride (3×10 mL ). Separate the aqueous phaseand freeze dry to give the title compound.

EXAMPLE 64s-[4α,7α(R*)12bβ]]-9-Amino-7-[(1-carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-6-oxopyrido[2,1-[[2]benzazepine-4-carboxylicacid dihydrochloride

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[(S-1-carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,87,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (260 mg, 385 mmol) in ethanol (7 mL). Add 10%palladium/carbon under an atmosphere of carbon dioxide. Shake under 42psi of hydrogen at room temperature for 5.5 hours. Filter and evaporatethe solvent in vacuo. Take up the residue in 1N hydrochloric acid (2 mL)and dilute with water (10 mL). Wash with methylene chloride (3×5 mL) andfreeze dry the aqueous phase to give the title compound.

EXAMPLE 7[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carbethoxy-3-phenylpropyl)amino[-1,2,3,4,6,7,8,12b-octahydro-1-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid

Scheme B, Step a:[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-amino-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (596 mg, 1.23 mmol) and proton sponge (296mg, 1.25 mmol) in methylene chloride (10 mL). Add a solution of(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid, ethyl ester (428mg, 1.25 mmol) in methylene chloride (6 ml). Stir at room temperatureunder an argon atmosphere overnight. Purify by silica gel chromatography(35% ethyl acetate/hexane) to give the title compound as a light yellowfoam (591 mg, 71.1%); mp 75°-80° C.

Anal. Calcd for C₄₀ H₄₁ N₃ O₇ : C, 71.09; H, 6.12; N, 6.22; Found: C,70.81; H, 5.98; N, 6.13.

Scheme B, Step b₁ :[4S-[4α,7α(R*),12bβ]]-7,[(S-1-Carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido]2,1-a]][2]benzazepine-4-carboxylicacid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[(S-1-carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (42.5 mg, 0.063 mmol) in methylene chloride(2 mL). Add trifluoroacetic acid (1 mL) and anisole (0.2 mL). Allow tostand for 2 hours under an argon atmosphere. Partition between methylenechloride and 9:1 water/methanol. Separate the organic phase and extractwith additional 9:1 water/methanol (10 mL). Combine the aqueous phasesand wash with methylene chloride (3×10 mL). Separate the aqueous phaseand freeze dry to give the title compound.

EXAMPLE 8[4S-[4α,7α(R*),12bβ]]11-Amino-7-[(S-1-carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, dihydrochloride

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[(S-1-carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid diphenylmethyl ester (260 mg, 38 5 mmol) in ethanol (7 mL). Add 10%palladium/carbon under an atmosphere of carbon dioxide. Shake under 42psi of hydrogen at room temperature for 5.5 hours. Filter and evaporatethe solvent in vacuo. Take up the residue in 1N hydrochloric acid (2 mL)and dilute with water (10 mL). Wash with methylene chloride (3×5 mL) andfreeze dry the aqueous phase to give the title compound as yellowcrystals (178 mg, 83.6%); mp 175°-95° C.

EXAMPLE 9[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino[3,4a][2]benzazepine-4-carboxylicacid

Scheme C, step a:N-[2-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-L-serine,methyl ester

Slurry N-phthaloyl-(S)-phenylalanine (90 g, 0.3 mol) in methylenechloride (450 mL) and add, by dropwise addition, oxalyl chloride (54 mL,0.62 mol). Place under a dry atmosphere (CaSO₄ tube) and treat withdimethylformamide (10 μL). Stir for 5 hours, filter and concentrate invacuo to give N-phthaloyl-(S)-phenylalanine, acid chloride as an offwhite amorphous solid.

Dissolve serine methyl ester hydrochloride (56 g, 0.36 mol) intetrahydrofuran (300 mL) then cool to 0° C. and add 4-methylmorpholine(88 mL, 0.8 mol). Add, by dropwise addition, a solution of theN-phthaloyl-(S)-phenylalanine, acid chloride in tetrahydrofuran (200mL). Allow to warm to room temperature and stir for 3 hours. Filter andconcentrate the filtrate in vacuo. Dissolve the residue in ethyl acetateand separate the organic phase. Wash with water then saturated sodiumchloride and dry (MgSO₄). Evaporate the solvent in vacuo to give an oil.Purify by silica gel chromatography (gradient 50% ethyl acetate/hexaneto ethyl acetate) to give the title compound (80.8 g, 67%) mp 129°-132°C.

Scheme C, step b:N-[2-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-O-2-propenyl-L-serine,methyl ester

DissolveN-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-L-serine,methyl ester (25 g, 63 mmol) in methylene chloride/cyclohexane (1:1, 600mL). Add allyl trichloroacetimidate (26 g, 128 mmol) andtrifluoromethanesulfonic acid (5 mL), 56.6 mmol). Stir at roomtemperature under a nitrogen atmosphere for 5 hours and dilute withmethylene chloride. Wash with saturated aqueous sodium hydrogencarbonate, water, dry (MgSO₄) and evaporate the solvent in vacuo. Purifyby silica gel chromatography (gradient 2 0% ethyl acetate/hexane to 35%ethyl acetate/hexane) to give the title compound; mp 95°-97° C.

Scheme C, step C: [S-(R*,R*)]-N-[2-(1,3-Dihydro-1,3-dioxo2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-3,4-dihydro-2H-1,4-oxazine-3-carboxylicacid, methyl ester

DissolveN-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-O-2-propenyl-L-serine,methyl ester (13 g, 29.8 mmol) in methylene chloride /methanol (10:1,220 mL). Cool to -78° C. and sparge with a mixture of ozone/oxygen forapproximately 10 minutes until a blue color persists. Sparge withnitrogen for 10 minutes at -78° C. to remove excess ozone. Treat withmethyl sulfide (60 mL, 0.82 mol) and allow to warm to room temperature.Stir at room temperature for 2.5 hours, evaporate the solvent in vacuoand dissolve the residue in ethyl acetate (200 mL). Wash with water,saturated sodium chloride, dry (MgSO₄) and evaporate the solvent invacuo to give the intermediateN-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-O-2-oxoethyl-L-serine,methyl ester as a foam (13.6 g).

DissolveN-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-O-2-oxoethyl-L-serine,methyl ester (13.6 g) in methylene chloride/trifluoroacetic acid(10:1/330 mL). Stir at room temperature for 2.5 hours and evaporate thesolvent in vacuo. Purify by silica gel chromatography (35% ethylacetate/hexane) and recrystallize (ethyl acetate/hexane) to give thetitle compound (8.52 g, 68%); mp 70°-72° C.

Scheme C, step d: [4S-[4α,7α(R*),12bβ]]-7-[(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)]-3,4,6,7,8,12b,hexahydro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylic acid

Dissolve [S-(R*,R*)]-N-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-3,4-dihydro-2H-1,4-oxazine-3-carboxylicacid, methyl ester (2.5 g, 5.9 mmol) in methylene chloride (5 mL) andadd, by dropwise addition, to a previously prepared solution oftrifluoromethanesulfonic acid (4.0 mL, 45 mmol) and trifluoroaceticanhydride (1.0 mL, 7.1 mmol). Place under a nitrogen atmosphere and stirat room temperature for 123 hours. Pour into a separatory funnelcontaining ice (200 g) and ethyl acetate (200 mL). Separate the organicphase, wash with water (3×200 mL) and saturated aqueous sodium chloride(100 mL). Extract the organic phase with 10% wt. potassium hydrogencarbonate (4×40 mL) and water (40 mL). Layer the combined basic aqueousphases with ethyl acetate (100 mL) and cool in an ice bath. Add, bydropwise addition, 6N hydrochloric acid to adjust the pH to 1 whilemaintaining the temperature at 5° -10° C. Separate the organic phase andextract the aqueous phase with ethyl acetate (3×200 mL ), wash withsaturated sodium chloride and dry (MgSO₄). Evaporate the solvent invacuo and dry the residue under high vacuum at 56° C. for 24 hours togive the title compound (1.75 g, 73%).

Scheme A, Step a:[4S-[4α,7α(R*),12bβ]]-7-[1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid and[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexaahydro-11-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid

Mix trifluromethanesulfonic acid (4 ml, 45 mmol) and nitric acid (1 ml,22 mmol) and anhydrous methylene chloride (100 mL). Cool to -78° C. andadd, by dropwise addition, a solution of[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid (1.94 g, 4.78 mmol) in methylene chloride (60 mL). Stir at -78° C.for 20 minutes, allow to warm to room temperature and stir under anitrogen atmosphere for 3.5 hours. Wash with water, dry (MgSO₄) andevaporate the solvent in vacuo to give yellow foam (2.3 g). Take up inethyl acetate, wash with brine (3×), dry (MgSO₄) and evaporate thesolvent in vacuo to give a yellow foam. Pufiy and separate by flashsilica gel chromatography 1:1:0.04 ethyl acetate/hexane/acetic acid) togive a small amount of the 9-nitro title compound and 1.02 g of the11-nitro title compound (47%).

Scheme A, Step b:[4S-[4α,7α(R*),12bβ]]-7-[1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylic acid, diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid (1.50 mmol) in methylene chloride (1 mL). Add diphenyldiazomethane(291 mg, 1.50 mmol) and let stand for several days in which time th emethylene chloride evaporates. Purify the residue by silica gelchromatography to give the title compound.

Scheme A, Step c:[4S-[4α,7α(R*),12bβ]]-7-Amino-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino-[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Slurry[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (600 mg, 0.97 mmol) in methanol (5 mL) andadd hydrazine hydrate (2.0 mL of a 1M solution in methanol, 2.0 mmol).Stir at room temperature under an argon atmosphere for 4 hours. Addmethanol (20 mL) and reflux for 1 hour, then stir at room temperatureovernight. Evaporate the solvent in vacuo, take up the residue in ethylacetate and filter. Wash with filtrate with water (3×) and brine. Dry(MgSO₄) and evaporate the solvent in vacuo to give the title compound(473 mg, 99.5%).

Scheme A, Step d:[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[Carbodiphenylmethoxy]-3-phenylpropyl]amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-amino-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (0. 632 mmol ) and proton sponge (138 mg,0.648 mmol) in methylene chloride (5 mL). Add a solution of(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid, diphenylmethylester (309 mg, 0.648 mmol) in methylene chloride (2 ml). Stir at roomtemperature under an argon atmosphere for 26 hours. Purify by flashsilica gel chromatography to give the title compound.

Scheme A, Step e₁ :[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[(diphenylmethoxy)carbonyl]-3-phenylpropyl]amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (0.045 mmol) in methylene chloride (1 mL).Add trifluoroacetic acid (1 mL) and anisole (0.2 mL). Allow to stand for3 hours under an argon atmosphere. Partition between methylene chlorideand 1N hydrochloric acid. Separate the aqueous phase and freeze dry.Purify by High Pressure Liquid Chromatography to give the titlecompound.

EXAMPLE 10 [4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid

Scheme A, Step a:[4S-[4α,7α(R*),12bβ]]-7-[1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4,-a][2]benzazepine-4-carboxylicacid and[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexaahydro-11-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, methyl ester (0.233 mmol) in ethanol (4 mL) and treat withtrifluoromethanesulfonic acid (0.5 mL). Stir under nitrogen atmosphereuntil hydrolysis is complete, pour into water, extract the aqueousphases with ethyl acetate (2×) and wash the combined organic phases withwater, then with saturated sodium chloride. Dry (MgSO₄), evaporate thesolvent in vacuo and purify by silica gel chromatography to give[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid.

Mix trifluromethanesulfonic acid (4 ml, 45 mmol) and nitric acid (1 ml,22 mmol ) and anhydrous methylene chloride (100 mL). Cool to -78° C. andadd, by dropwise addition, a solution of[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid (4.78 mmol) in methylene chloride (60 mL). Stir at -78° C. for 20minutes, allow to warm to room temperature and stir under a nitrogenatmosphere for 3.5 hours. Wash with water, dry (MgSO₄) and evaporate thesolvent in vacuo. Purify and separate by flash silica gel chromatographyto give the 11-nitro title compound and the 9-nitro title compound.

Scheme A, Step b:[4S-[4α,7α(R*),12bβ]]-7-[1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid (1.50 mmol) in methylene chloride (1 mL). Add diphenyldiazomethane(291 mg, 1.50 mmol) and let stand for several days in which time themethylene chloride evaporates. Purify the residue by silica gelchromatography to give the title compound.

Scheme A, Step C:[4S-[4α,7α(R*),12bβ]]-7-Amino-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester

Slurry[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylic acid, diphenylmethyl ester (0.324 mmol) inmethanol (6 mL) and add hydrazine hydrate (1.0 mL of a 1M solution inmethanol, 1.0 mmol). Stir at room temperature under an argon atmospherefor 20 hours. Dilute with methylene chloride (30 mL) and stir for 2hours. Filter and concentrate in vacuo. Take up the residue in methylenechloride and filter again. Wash the filtrate with water, dry (MgSO₄) andevaporate the solvent in vacuo to give the title compound.

Scheme A, Step d:[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[Carbodiphenylmethoxy]-3-phenylpropyl]amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylic acid, diphenyl methyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-amino-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (0.632 mmol) and proton sponge (138 mg, 0.648mmol) in methylene chloride (5 mL). Add a solution of(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid, diphenylmethylester (309 mg, 0.648 mmol) in methylene chloride (2 ml). Stir at roomtemperature under an argon atmosphere for 26 hours. Purify by flashsilica gel chromatography to give the title compound.

Scheme A, Step e₁ :4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[[S-1-[carbodiphenylmethoxy]-3-phenylpropyl]amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, diphenylmethyl ester (0.045 mmol) in methylene chloride (1 mL).Add trifluoroacetic acid (1 mL) and anisole (0.2 mL). Allow to stand for3 hours under an argon atmosphere. Partition between methylene chlorideand 1N hydrochloric acid. Separate the aqueous phase and freeze dry.Purify by High Pressure Liquid Chromatography to give the titlecompound.

EXAMPLE 11[4S-[4α,7α(R*),12bβ]],7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-][2]benzazepine-4-carboxylic acid

Scheme D, step a:N-[2-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2,yl)-1-oxo-3-phenylpropyl]-(S)-3-[(trifluoroacetyl-2-propenyl)amino]-2-amino-propionicacid, methyl ester

Dissolve N.sup.α -(carbobenzyloxy)-β-(amino)-L-alanine, methyl ester(2.27 mmol) in anhydrous tetrahydrofuran (15 mL). Treat with pyridine(183 μL, 2.27 mmol) followed by trifluoroacetic anhydride (321 μL, 2.27mmol) and stir at ambient temperature for 1 hours. Add additionalpyridine (180 μL) and trifluoroacetic anhydride (320 μL). Allow to standovernight, partition between ethyl ether and water. Separate the organicphase, dry (MgSO₄) and evaporate the solvent in vacuo to give N.sup.α-(carbobenzyloxy)-β-(trifluoroacetyl)-L-alanine, methyl ester.

Suspend sodium hydride (4.8 g, 0.2 mol) in anhydrous dimethylformamide(100 mL), cool to 0° C. and place under a nitrogen atmosphere. Add, bydropwise addition, a solution of N.sup.α-(carbobenzyloxy)-β-(trifluoroacetyl)-L-alanine, methyl ester (0.2 mol)in dimethylformamide. Stir until evolution of hydrogen ceases. Add, bydropwise addition, a solution of allyl bromide (0.2 mol) indimethylformamide (100 mL). Stir overnight at room temperature thencarefully quench with saturated ammonium chloride. Extract into ethylacetate, dry (MgSO₄) and evaporate the solvent in vacuo. Purify bysilica gel chromatography to give N.sup.α-(carbobenzyloxy)-β-(trifluoroacetyl-allylamino)-L-alanine, methylester.

Place boron tribromide (215 mg, 0.86 mmol) in a flask and cool to 0° C.Cautiously add trifluoroacetic acid (5 mL) with stirring. Evaporate thesolvent to give boron tris(trifluoroacetate).

Dissolve boron tris(trifluoroacetate) (0.3 g, 0.86 mmol) intrifluoroacetic acid (10 mL) and add N.sup.α-(carbobenzyloxy)-β-(trifluoroacetyl-allylamino)-L-alanine, methyl ester(105 mg, 0.27 mmol). Stir under an argon atmosphere for 1 hour thenevaporate the solvent in vacuo at room temperature. Add methanol andevaporate repeatedly. Purify by silica gel chromatography to giveβ-(trifluoroacetyl-allylamino)-L-alanine, methyl ester, hydrochloride.

Dissolve β-(trifluoroacetyl-allylamino)-L-alanine, methyl ester,hydrochloride (104.8 g, 0.36 mol) in tetrahydrofuran (300 mL) then coolto 0° C. and add 4-methylmorpholine (88 mL, 0.8 mol). Add, by dropwiseaddition, a solution of the N-phthaloyl-(S)-phenylalanine, acid chloride(108.7 g, 0.36 mol) in tetrahydrofuran (200 mL). Allow to warm to roomtemperature and stir for 3 hours. Filter and concentrate the filtrate invacuo. Dissolve the residue in ethyl acetate and separate the organicphase. Wash with water then saturated sodium chloride and dry (MgSO₄).Evaporate the solvent in vacuo to give an oil. Purify by silica gelchromatography to give the title compound.

Scheme D, step b: [S-(R*,R*)]-N-[2-(1.3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-3,4-dihydro-2H-4-trifluoroacetyl-1,4-azazine-3-carboxylicacid, methyl ester

Dissolve N-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-(S)-3-[(trifluoroacetyl-2propenyl)amino]-2-amino-propionic acid,methyl ester (15.8 g, 29.8 mmol) in methylene chloride/methanol (10:1,220 mL). Cool to -78° C. and sparge with a mixture of ozone/oxygen forapproximately 10 minutes until a blue color persists. Sparge withnitrogen for 10 minutes at -78° C. to remove excess ozone. Treat withmethyl sulfide (60 mL, 0.82 mol) and allow to warm to room temperature.Stir at room temperature for 2.5 hours, evaporate the solvent in vacuoand dissolve the residue in ethyl acetate (200 mL). Wash with water,saturated sodium chloride, dry (MgSO₄) and evaporate the solvent invacuo to give the intermediateN-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl-]-(S)-3[(trifluoroacetyl-2-oxoethyl)amino]-2-amino-propionicacid, methyl ester.

DissolveN-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-(S)-3-[(trifluoroacetyl-2-oxoethyl)amino-propionicacid, methyl ester (15.9 g, 29.8 mmol) in methylenechloride/trifluoroacetic acid (10:1/330 mL). Stir at room temperaturefor 2.5 hours and evaporate the solvent in vacuo. Purify by silica gelchromatography to give the title compound.

Scheme D, step c: [4S-[4α,7α(R*),12bβ]]-7-[(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)]-3,4,6,7,8,12b-hexahydro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid

Dissolve [S-(R*,R*)]-N-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-1-oxo-3-phenylpropyl]-3,4-dihydro-2H-4-trifluoracetyl-1,4-azazine-3-carboxylicacid, methyl ester (3.04 g, 5.9 mmol) in methylene chloride (5 mL) andadd, by dropwise addition, to a previously prepared solution oftrifluoromethanesulfonic acid (4.0 mL, 45 mmol) and trifluoroaceticanhydride (1.0 mL, 7.1 mmol). Place under a nitrogen atmosphere and stirat room temperature for 123 hours. Pour into a separatory funnelcontaining ice (200 g) and ethyl acetate (200 mL). Separate the organicphase, wash with water (3×200 mL) and saturated aqueous sodium chloride(100 mL). Extract the organic phase with 10% wt. potassium hydrogencarbonate (4×40 mL) and water (40 mL). Layer the combined basic aqueousphases with ethyl acetate (100 mL) and cool in an ice bath. Add, bydropwise addition, 6N hydrochloric acid to adjust the pH to 1 whilemaintaining the temperature at 5°-10° C. Separate the organic phase andextract the aqueous phase with ethyl acetate (3×200 mL), wash withsaturated sodium chloride arid dry (MgSO₄). Evaporate the solvent invacuo and dry the residue under high vacuum at 56° C. for 24 hours togive the title compound.

Scheme A, step a:[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid and[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid (3.5mmol) in methylene chloride (10 mL) and cool to -60° C. Add, by dropwiseaddition, a solution of nitronium tetrafluoroborate (25 mL of a 0.5Msolution in sulfolane, 12.5 mmol) in methylene chloride (15 mL). Warmslowly to 10° C. over 34 hours. Partition between methylene chloride (75mL) and water (75 mL). Separate the aqueous phase and extract withmethylene chloride (50 mL). Combine the organic phases, dry (MgSO₄) andevaporate the solvent in vacuo. Purify and separate by flash silica gelchromatography to give the 11-nitro title compound and the 9-nitro titlecompound.

Scheme A, Step b:[4S-[4α,7α(R*),12bβ]]-7-[1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid (1.50mmol) in methylene chloride (1 mL). Add diphenyldiazomethane (291 mg,1.50 mmol) and let stand for several days in which time the methylenechloride evaporates. Purify the residue by silica gel chromatography togive the title compound.

Scheme A, Step c:[4S-[4α,7α(R*),12bβ]]-7-Amino-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acids.diphenylmethyl ester

Slurry[4S-[4α,7α(R*),12bβ]]-7-[1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,diphenylmethyl ester (0. 324 mmol) in methanol (6 mL) and add hydrazinehydrate (1.0 mL of a 1M solution in methanol, 1.0 mmol). Stir at roomtemperature under an argon atmosphere for 20 hours. Dilute withmethylene chloride (30 mL) and stir for 2 hours. Filter and concentratein vacuo. Take up the residue in methylene chloride and filter again.Wash the filtrate with water, dry (MgSO₄) and evaporate the solvent invacuo to give the title compound.

Scheme A, Step d:[4S-[4α,7α(R*),12bβ]]-7-[[S-1[Carbodiphenylmethoxy]-3-phenylpropyl]amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,diphenylmethyl ester

Dissolve[4S-[4α,7α(R*),12bβ]]-7-amino-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,diphenylmethyl ester (0.632 mmol) and proton sponge (138 mg, 0.648 mmol)in methylene chloride (5 mL). Add a solution of(R)-2-trifluoromethanesulfonyl-4-phenylbutyric acid, diphenylmethylester (309 mg, 0.648 mmol) in methylene chloride (2 ml). Stir at roomtemperature under an argon atmosphere for 26 hours. Purify by flashsilica gel chromatography to give the title compound.

Scheme A, Step e₁ :[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid

Dissolve[4S-[4α,7α(R*),12bβ]]07-[[S-1-[carbodiphenylmethoxy]-3-phenylpropyl]amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,diphenylmethyl ester (0.045 mmol) in methylene chloride (1 mL). Addtrifluoroacetic acid (1 mL) and anisole (0.2 mL). Allow to stand for 3hours under an argon atmosphere. Partition between methylene chlorideand 1N hydrochloric acid. Separate the aqueous phase and freeze dry.Purify by High Pressure Liquid Chromatography to give the titlecompound.

EXAMPLE 12[4S,[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-azazino[3,4-a][2]benzazepine-4-carboxylicacid

Dissolve[4S-[4α,7α(R*),12bβ]]-7-[(S-1-carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid (2.3mmol) in tetrahydrofuran (10 mL) at 0° C. then treat with 1N lithiumhydroxide (6.75 mL). Add ethanol(2-3 mL) and stir for 30 minutes at 0°C. Partition between ethyl acetate and water and separate the organicphase. Wash with water, dry (MgSO₄) and evaporate the solvent in vacuoto give the title compound.

The following compounds can be prepared by analogous procedures to thosedescribed above in Examples 1-12:

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-amino-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-amino-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-amino-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-amino-6-oxo-1H-[1,4]-N⁴-trifluoroacetyl-azazino[3,4-a][2]benzazepine-4-carboxylic acid,dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-azazino[3,4-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-amino-6-oxo-1H-[1,4]-azazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-amino-6-oxo-1H-[1,4]-azazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-azazino[3,4-a][2]benzazepine-4-carboxylic acid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-azazino[3,4-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-amino-6-oxo-1H-[1,4]-azazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-amino-6-oxo-1H-[1,4]-azazino[3,4-a][2]benzazepine-4-carboxylic acid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-amino-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-amino-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid;

4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-amino-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-amino-6-oxo-1H-[1,4]-thiazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-amino-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-amino-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-nitro-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-9-amino-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboethoxy-3-phenylpropyl)amino]-3,4,6,7,8,12b-hexahydro-11-amino-6-oxo-1H-[1,4]-oxazino[3,4-a][2]benzazepine-4-carboxylic acid, dihydrochloride.

In a further embodiment, the present invention provides a method ofinhibiting ACE in a patient in need thereof comprising administering tosaid patient an effective ACE inhibitory amount of a compound of formula(1).

As used herein, the term "patient" refers to warm-blooded animals ormammals, including mice, rats and humans. A patient is in need oftreatment to inhibit ACE when it would be beneficial to the patient toreduce the physiological effect of circulating angiotensin II. Forexample, a patient is in need of treatment to inhibit ACE when thepatient is suffering from hypertension, chronic congestive heartfailure, hyperaldosteronemia or cognitive disorders. Inhibition of ACEreduces levels of angiotensin II and thus inhibits the vasopressor,hypertensive and hyper-aldosteronemic effects caused thereby.

An effective ACE inhibitory amount of a compound of formula (1) is thatamount which is effective in inhibiting ACE in a patient in need thereofwhich results, for example, in a hypotensive effect.

An effective ACE inhibitory dose can be readily determined by the use ofconventional techniques and by observing results obtained underanalogous circumstances. In determining the effective dose, a number offactors are considered including, but not limited to: the species ofpatient; its size, age, and general health; the specific diseaseinvolved; the degree of or involvement or the severity of the disease;the response of the individual patient; the particular compoundadministered; the mode of administration; the bioavailabilitycharacteristics of the preparation administered; the dose regimenselected; and the use of concomitant medication.

An effective ACE inhibitory amount of a compound of formula (1) willgenerally vary from about 0.01 milligram per kilogram of body weight perday (mg/kg/day) to about 20 mg/kg/day. A daily dose of from about 0.1mg/kg to about 10 mg/kg is preferred.

In effecting treatment of a patient, compounds of formula (1) can beadministered in any form or mode which makes the compound bioavailablein effective amounts, including oral and parenteral routes. For example,the compound can be administered orally, subcutaneously,intramuscularly, intravenously, transdermally, intranasally, rectally,and the like. Oral administration is generally preferred. One skilled inthe art of preparing formulations can readily select the proper form andmode of administration depending upon the disease state to be treated,the stage of the disease, and other relevant circumstances.

Compounds of formula (1) can be administered in the form ofpharmaceutical compositions or medicaments which are made by combiningthe compounds of formula (1) with pharmaceutically acceptable carriersor excipients, the proportion and nature of which are determined by thechosen route of administration, and standard pharmaceutical practice.

In another embodiment, the present invention provides compositionscomprising a compound of formula (1) in admixture or otherwise inassociation with one or more inert carriers. These compositions areuseful, for example, as assay standards, as convenient means of makingbulk shipments, or as pharmaceutical compositions. An assayable amountof a compound of formula (1) is an amount which is readily measurable bystandard assay procedures and techniques as are well known andappreciated by those skilled in the art. Assayable amounts of a compoundof formula (1) will generally vary from about 0.001% to about 75% of thecomposition by weight. Inert carriers can be any material which does notdegrade or otherwise covalently react with a compound of formula (1).Examples of suitable inert carriers are water; aqueous buffers, such asthose which are generally useful in High Performance LiquidChromatography (HPLC) analysis; organic solvents, such as acetonitrile,ethyl acetate, hexane and the like; and pharmaceutically acceptablecarriers or excipients.

More particularly, the present invention provides pharmaceuticalcompositions comprising an effective amount of a compound of formula (1)in admixture or otherwise in association with one or morepharmaceutically acceptable carriers or excipients.

The pharmaceutical compositions or medicaments are prepared in a mannerwell known in the pharmaceutical art. The carrier or excipient may be asolid, semi-solid, or liquid material which can serve as a vehicle ormedium for the active ingredient. Suitable carriers or excipients arewell known in the art. The pharmaceutical composition may be adapted fororal or parenteral use and may be administered to the patient in theform of tablets, capsules, suppositories, solution, suspensions, or thelike.

The pharmaceutical compositions may be administered orally, for example,with an inert diluent or with an edible carrier. They may be enclosed ingelatin capsules or compressed into tablets. For the purpose of oraltherapeutic administration, the compounds of formula (1) may beincorporated with excipients and used in the form of tablets, troches,capsules, elixirs, suspensions, syrups, wafers, chewing gums and thelike. These preparations should contain at least 4% of the compound offormula (1), the active ingredient, but may be varied depending upon theparticular form and may conveniently be between 4% to about 70% of theweight of the unit. The amount of the active ingredient present incompositions is such that a unit dosage form suitable for administrationwill be obtained.

The tablets, pills, capsules, troches and the like may also contain oneor more of the following adjuvants: binders, such as microcrystallinecellulose, gum tragacanth or gelatin; excipients, such as starch orlactose, disintegrating agents such as alginic acid, Primogel, cornstarch and the like; lubricants, such as magnesium stearate or Sterotex;glidants, such as colloidal silicon dioxide; and sweetening agents, suchas sucrose or saccharin may be added or flavoring agents, such aspeppermint, methyl salicylate or orange flavoring. When the dosage unitform is a capsule, it may contain, in addition to materials of the abovetype, a liquid carrier such as polyethylene glycol or a fatty oil. Otherdosage unit forms may contain other various materials which modify thephysical form of the dosage unit, for example, as coatings. Thus,tablets or pills may be coated with sugar, shellac, or other entericcoating agents. A syrup may contain, in addition to the activeingredient, sucrose as a sweetening agent and certain preservatives,dyes and colorings and flavors. Materials used in preparing thesevarious compositions should be pharmaceutically pure and non-toxic inthe amounts used.

For the purpose of parenteral administration, the compounds of formula(1) may be incorporated into a solution or suspension. Thesepreparations should contain at least 0.1% of a compound of theinvention, but may be varied to be between 0.1 and about 50% of theweight thereof. The amount of the active ingredient present in suchcompositions is such that a suitable dosage will be obtained.

The solutions or suspensions may also include one or more of thefollowing adjuvants: sterile diluents such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl paraben; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylene diaminetetraacetic acid;buffers such as acetates, citrates or phosphates and agents for theadjustment of toxicity such as sodium chloride or dextrose. Theparenteral preparation can be enclosed in ampules, disposable syringesor multiple dose vials made of glass or plastic.

As with any group of structurally related compounds which possess aparticular generic utility, certain groups and configurations arepreferred for compounds of formula (1) in their end-use application.

The compounds of formula (1) wherein X is amino and Y is hydrogen aregenerally preferred.

It is, of course, understood that the compounds of formula (1) exist inparticular isomeric configurations including structural as well asstereo isomers. It is further understood that the present inventionencompasses those compounds in the particular isomeric configurationsdepicted by the structure of formula (1) which utilizes standardtechniques and conventions for drawing isomeric structures.

The following specific compounds of formula (1) are particularlypreferred in the end-use application of the compounds of the presentinvention:

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-9-Amino-7-[(S-1-carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-6oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-11-Amino-7-[(S-1-carboxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-6oxopyrido[2,1-a][2]benzazepine-4-carboxylic acid, dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-9-nitro-6oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-9-Amino-7-[(1-carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-6-oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid dihydrochloride;

[4S-[4α,7α(R*),12bβ]]-7-[(S-1-Carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-11-nitro-6oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid;

[4S-[4α,7α(R*),12bβ]]-11-Amino-7-[(S-1-carbethoxy-3-phenylpropyl)amino]-1,2,3,4,6,7,8,12b-octahydro-6oxopyrido[2,1-a][2]benzazepine-4-carboxylicacid, dihydrochloride;

The utility of the compounds of formula (1) may be demonstrated bymonitoring the inhibition of ACE in vitro using the spectrophotometricsubstrate described by Holmquist et al. [Anal. Biochem. 95, 540-548(1979)] and the buffer system described by Ryan [Methods of EnzymaticAnalysis, 3rd ed., H. U. Bergmeyer, editor; vol. V, Verlag Chemie,Weinheim, 1983, pp. 20-34].

The amino or nitro substituted compounds of the present inventionpossess an unexpectedly prolonged duration of activity as compared toother ACE inhibitors similar in structure.

What is claimed is:
 1. A compound of the formula ##STR7## wherein A issulfur;R is hydrogen or a C₁ -C₄ alkyl; and X and Y are eachindependently hydrogen, nitro or amino, with the proviso that one of Xand Y is hydrogen and one of X and Y is other than hydrogen;andpharmaceutically acceptable salts thereof.
 2. A compound of claim 1wherein Y is nitro and X is hydrogen.
 3. A compound of claim 1 wherein Yis amino and X is hydrogen.
 4. A compound of claim 1 wherein X is nitroand Y is hydrogen.
 5. A compound of claim 1 wherein X is amino and Y ishydrogen.
 6. A pharmaceutical composition comprising an effective ACEinhibitory amount of a compound of claim 1 in admixture or otherwise inassociation with one or more pharmaceutically acceptable carriers orexcipients.
 7. A method of inhibiting ACE in a patient in need thereofcomprising administering to said patient an effective ACE inhibitoryamount of a compound of claim
 1. 8. A compound of claim 1 wherein A issulfur.
 9. A method of claim 7 wherein the patient is suffering fromhypertension.
 10. A method of claim 7 wherein the patient is sufferingfrom chronic congestive heart failure.